In this study we characterised the development of caecal microbiota in egg laying hens over their commercial production lifespan, from the day of hatching until 60 weeks of age. Using pyrosequencing ...of V3/V4 variable regions of 16S rRNA genes for microbiota characterisation, we were able to define 4 different stages of caecal microbiota development. The first stage lasted for the first week of life and was characterised by a high prevalence of Enterobacteriaceae (phylum Proteobacteria). The second stage lasted from week 2 to week 4 and was characterised by nearly an absolute dominance of Lachnospiraceae and Ruminococcaceae (both phylum Firmicutes). The third stage lasted from month 2 to month 6 and was characterised by the succession of Firmicutes at the expense of Bacteroidetes. The fourth stage was typical for adult hens in full egg production aged 7 months or more and was characterised by a constant ratio of Bacteroidetes and Firmicutes formed by equal numbers of the representatives of both phyla.
The olm (Proteus anguinus) is a troglomorphic, neotenous amphibian with extraordinary life expectancy and unique adaptations that deserve further investigation. A low reproductive rate and habitat ...decline render it threatened by extinction. Establishing captive populations for maintenance and artificial breeding may one day become crucial to the species. Longitudinal, in-vivo assessment of inner organs is invaluable to our understanding of reproductive physiology, health, and behavior. Using ultrasound, we measured heart rate and assessed health and reproductive status of 13 captive olms at Zagreb Zoo. Heart rate averaged 42.9 ± 4.6 bpm (32-55 bpm), as determined via pulsed-wave Doppler at 4-12 MHz. By using frequencies of up to 70 MHz (ultrasound biomicroscopy), inner organs were visualized in detail. Assessment of the gastrointestinal tract provided insights into feeding status and digestive processes. Several subclinical pathologies were detected, including biliary sludge, subcutaneous edema, ascites, and skin lesions. Detection of skin lesions by ultrasound was more sensitive than visual adspection. Olms with ultrasonographically detected skin lesions tested positive for Saprolegnia and were treated. Three of the four affected individuals survived and subsequently tested negative for Saprolegnia. Sex was reliably determined; only one individual proved male. The reason for this extreme female-biased sex-ratio remains unknown. However, as most of the individuals were flushed from the caves by strong currents in spring, the sample may not be representative of natural populations. In female olms, different stages of ovarian follicular development were observed with diameters ranging between 0.1 and 1.1 mm. Results were confirmed by comparing ultrasound, necropsy, and histological findings of one dead specimen. In summary, ultrasound proved a valuable tool to support conservation and captive breeding programs by allowing non-invasive assessment of physiological parameters, clinical condition, and reproductive status in olms.
Mycoplasma synoviae (MS), despite its lower pathogenicity, has the ability to cause significant losses in poultry production but is usually underdiagnosed. In egg layer production, losses could be ...significant because of the drop in egg production and poor eggshell quality. Problems with colibacillosis and other infections secondary to MS have been continuously reported on Croatian egg layer farms. As a result, regular monitoring of flocks also included screening of the seroprevalence and molecular detection of MS using ELISA and qPCR tests, respectively. During monitoring, altogether 1135 samples were tested, including 1067 serum samples and 68 tracheal swabs, in a total of 126 flocks and 83 longitudinally merged flocks on 15 farms during the period from 2017 to 2021. The results showed a high general prevalence of MS with 86.6% positive layer farms, while grouped flock seroprevalence and prevalence were 98.6% and 85.7%, respectively. With age, seroprevalence and ELISA titers rise significantly compared to the rearing period, with a significant mutual correlation over the entire production period. Additionally, there is a significant correlation between ELISA titers and age in weeks. Several flocks covered longitudinally from the first week of age, over the rearing period to the end of production, showed low prevalence during the rearing period, with a later significant rise in titer and prevalence, which indicates the dominance of horizontal transmission during production. Overall results indicate the need for a prompt reaction regarding preventive measures, such as better flock management, biosecurity and vaccination, which would reduce the losses and improve production. Key words: Mycoplasma synoviae; layer flocks; seroprevalence; ELISA; molecular detection; qPCR Mycoplasma synoviae (MS), unatoc nizoj patogenosti, moze uzrokovati znacajne gubitke u proizvodnji peradi, no obicno je poddijagnosticirana. Gubici u proizvodnji konzumnih jaja mogu biti znacajni zbog pada proizvodnje i lose kvalitete ljuske jajeta. Problemi s kolibacilozom i drugim zarazama koje se javljaju kao sekundarne MS-u kontinuirano se prijavljuju na farmama nesilica konzumnih jaja u Hrvatskoj. Stoga je u pracenje zdravlja na farmama ukljuceno i otkrivanje seroprevalencije i molekularnog dokaza MS-a primjenom ELISA i qPCR testova. Tijekom pracenja testirano je ukupno 1135 uzoraka, 1067 seruma i 68 obrisaka dusnika, u ukupno 126 jata, odnosno 83 longitudinalno grupiranih jata na 15 farmi od 2017. do 2021. godine. Rezultati su pokazali visoku opcu prevalenciju od 86,6% pozitivnih farmi na MS. Kod longitudinalno grupiranih jata seroprevalencija je iznosila 98,6%, dok je molekularnim monitoringom prevalencija bila 85,7%. S dobi, seroprevalencija i ELISA titar znacajno rastu, uz znacajnu medusobnu korelaciju tijekom cijelog razdoblja proizvodnje. Takoder, postoji znacajna korelacija izmedu ELISA titra i dobi u tjednima. Jata pracena longitudinalno od dobi jednog tjedna do kraja proizvodnje, pokazuju nisku prevalenciju tijekom uzgojnog razdoblja. Kasnije, u razdoblju proizvodnje, dolazi do znacajnog porasta titra i prevalencije, sto ukazuje na dominantan horizontalni prijenos tijekom proizvodnje. Rezultati monitoringa skrecu pozornost na potrebu brze reakcije na farmama s ciljem poboljsanja preventivnih mjera, narocito boljeg upravljanja jatima, biosigurnosnih mjera i cijepljenja, sto bi znacajno smanjilo gubitke i poboljsalo proizvodnju. Kljucne rijeci: Mycoplasma synoviae; kokosi nesilice; seroprevalencija; ELISA; prevalencija; qPCR
Proteus anguinus (olm) is an amphibian endemic to underground cave systems of the Dinaric karst of Central Europe, adapted to a life in complete darkness. Recent studies have demonstrated the global ...decline of amphibians due to poorly understood microbiological diseases, emphasizing just how little is known about the natural microflora of amphibians. Not much is known about the olm’s microbiota in their cave habitats, and nothing is known about the microbiology of animals washed to the surface. This observational study describes the microbiological analysis of the skin, cloaca and oral cavity of six olms collected from their cave habitats, and 16 collected after being washed out by flows from two springs, to learn more about the olm’s normal microbiota and possible changes after contact with the different environmental conditions on the surface. Standard microbiological procedures, MALDI-TOF and Real-Time PCR were used for microbiological species identification. All animals tested negative for Ranavirus, Batrachochytrium dendrobatidis, Batrachochytrium salamandrivorans and Chlamydia spp. The most abundant fungi isolated were Penicillium spp. and Cladosporium spp. The washed out animals showed a higher diversity of bacterial flora than those from cave habitats, with Acinetobacter johnsonii, Aeromonas hydrophila, Bacillus spp. and Janthinobacterium lividum as the most frequently identified isolates.
The Martino’s vole (
Dinaromys bogdanovi
) is a rare species that lives in differently structured karst habitats, varying from open rocky plains to deep fissures, mostly residing under boulders and ...in crevices. Populations of the species are declining, probably due to its strict habitat preferences and competition with the European snow vole. Since the species is difficult to study in the wild, we tested its behavioral response to a differently complex environment and novel object presence in captivity. We exposed 14 individuals to differentially complex setups, from open and unsheltered to rocky with covered tunnels. We measured the effect of the presented setup and season on vole behavior. Analysis showed seasonal differences with higher movement frequency and longer time spent still during the breeding season, and a higher number of attempts due to the lack of vole entry into the experimental terrarium in the non-breeding season. Additionally, movement significantly differentiated between an open and simple setup in comparison to more complex ones, with higher frequencies of moving and peeping and the duration of peeping in the simple setup, indicating restlessness. We also found significant differences between the novel object and control setup. The results of this study could be useful for future assessment of the influence of habitat complexity on mobility in the wild. Moreover, this is the first study on the behavior of this endemic and rare rodent.
Emerging infections add to existing threats to the survival of amphibians worldwide. The olm (Proteus anguinus) is a vulnerable, troglobiont urodele species with a small European range and restricted ...to underground karstic systems. Population declines to emerging threats like the chytrid fungus Batrachochytrium salamandrivorans, are likely to go unnoticed due to inaccessibility of the species' habitat. We here studied the interaction between olms and B. salamandrivorans. Experimental inoculation of olms resulted in low-level, asymptomatic but persistent infections, with limbs as predilection sites. The lack of exponential fungal growth in the olms' epidermis correlated with limited fungal proliferation and dampened virulence gene expression after exposure to olm skin compounds. The olm is one of few western Palearctic urodeles that is tolerant to B. salamandrivorans infection and may act as a subterranean disease reservoir, yet costs of subclinical infection may compromise olm fitness on the long term.
Tapeworms of the genus
Mesocestoides
(Cestoda: Cyclophyllidea: Mesocestoididae) are still enigmatic to scientists, due to their high morphological variability, low host specificity, and unknown ...details of their life cycle. They are found worldwide, with carnivorous mammals as the main definitive hosts, and the disease is potentially zoonotic. After ingestion by a definitive host, the tetrathyridium can occasionally migrate through the intestinal wall and reach the peritoneal cavity or abdominal organs causing peritoneal metacestodosis. Here, we report on a case of metacestodosis of a European wild cat (
Felis silvestris silvestris
) found dead in Croatia. At necropsy, a large number of white, rice-like structures were found free in the abdominal and thoracic cavities, as well as along the serous surfaces and in the lungs. DNA isolated from the nodules was genotyped and based on a 320-base pair long 12S fragment classified as
Mesocestoides vogae
. Although post-mortem changes were advanced, severe emaciation due to the severe parasitic infection and gastrointestinal bleeding was diagnosed as the likely cause of death. Intestinal cestodosis was previously reported in wild cats, but according to our knowledge, this is the first description of peritoneal and pleural metacestodosis caused by
M. vogae
tetrathyridia (metacestodes) in any wild carnivore species.
Salmonellosis transmitted by pet reptiles is an increasing public health issue worldwide. The aim of this study was to investigate the prevalence of Salmonella strains from captive reptiles in ...Croatia. From November 2009 to November 2011 a total of 292 skin, pharyngeal, cloacal, and fecal samples from 200 apparently healthy reptiles were tested for Salmonella excretions by bacteriologic culture and serotyping. These 200 individual reptiles included 31 lizards, 79 chelonians, and 90 snakes belonging to private owners or housed at the Zagreb Zoo, Croatia. Salmonella was detected in a total of 13% of the animals, among them 48.4% lizards, 8.9% snakes, and 3.8% turtles. Representatives of five of the six Salmonella enterica subspecies were identified with the following proportions in the total number of isolates: Salmonella enterica enterica 34.6%, Salmonella enterica houtenae 23.1%, Salmonella enterica arizonae 23.1%, Salmonella enterica diarizonae 15.4%, and Salmonella enterica salamae 3.8%. The 14 different serovars isolated included several rarely occurring serovars such as Salmonella Apapa, Salmonella Halle, Salmonella Kisarawe, and Salmonella Potengi. These findings confirm that the prevalence of Salmonella is considerable in captive reptiles in Croatia, indicating that these animals may harbor serovars not commonly seen in veterinary or human microbiologic practice. This should be addressed in the prevention and diagnostics of human reptile-transmitted infections.
PDF
